Identification and characterization of Helicobacter pylori phospholipase C activity

Jörn Hendrik Weitkamp, Guillermo I. Pérez-Pérez, Günter Bode, Peter Malfertheiner, Martin J. Blaser

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

We analyzed 11 H. pylori isolates from humans using the artificial chromogenic substrate paranitrophenylphosphorylcholine to detect phospholipase C (PLC) activity. The range of PLC in sonicates was 8.8–92.3 (Mean 56.9 ± 6.5) nmol of substrate hydrolysed min−1 mg−1 protein; the amount of activity was not associated with urease or cytotoxin levels. Addition of sorbitol or glycerol enhanced PLC activity of H. pylori sonicate and purified PLC from C. perfringens (PLC 1) but not purified PLC from B. cereus (PLC3 ). H. pylori sonicates had little acid phosphatase and no detectable alkaline phosphatase activity, and H. pylori PLC showed markedly different biochemical characteristics from either phosphatase. In total, these studies indicate that activity measured in H. pylori sonicate by PLC assay is due to PLC and not phosphatase activity. The temperature optimum for PLC activity of H. pylori sonicate was 56 °C and for PLC 1 was 65 °C. For H. pylori PLC and PLC 1, optimal activity occurred at pH 8. Despite multiple similarities between H. pylori PLC and PLC1, known PLC inhibitors show different interactions with each enzyme. Although PLC activity is present in many subcellular constituents of H. pylori, including culture supernatants and water extracts, highest specific activity is associated with a membrane-enriched fraction.

Original languageEnglish (US)
Pages (from-to)11-27
Number of pages17
JournalZentralblatt fur Bakteriologie
Volume280
Issue number1-2
DOIs
StatePublished - 1993
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Immunology

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